Results: 5

1.
Fig. 2

Fig. 2. From: EBV-positive human sera contain antibodies against the EBV BMRF-2 protein.

Detection of BMRF-2 in Akata/BMRF-2 and 293T/BMRF-2 cells using human serum samples. Akata/BMRF-2 (A) and 293T/BMRF-2 (B) cells were immunostained with EBV-positive NPC-4 (A and B, upper panels) and EBV-negative human sera (Wampole Laboratories) (A and B, lower panels). BMRF-2 is shown in green, human serum signals are red, and yellow in merged panels indicates co-localization of the BMRF-2 GFP signal with human sera-specific red signal. Cell nuclei were stained in blue.

Jianqiao Xiao, et al. Virology. ;393(1):151-159.
2.
Fig. 3

Fig. 3. From: EBV-positive human sera contain antibodies against the EBV BMRF-2 protein.

Detection of cell surface BMRF-2 by flow cytometry using human serum samples. Akata/BMRF-2 and 293T/BMRF-2 cells were incubated with human sera and RPE-labeled goat anti-human IgG was used to detect binding of human anti-BMRF-2 antibodies to the cell surface. Gray-filled curves represent EBV-negative human serum (Wampole Laboratories) and unfilled curves indicate EBV-positive serum samples. MN, mean fluorescence intensity.

Jianqiao Xiao, et al. Virology. ;393(1):151-159.
3.
Fig. 4

Fig. 4. From: EBV-positive human sera contain antibodies against the EBV BMRF-2 protein.

Western blot analysis of human sera reactivity to the BMRF-2 RGD and mutant BMRF-2 AAA peptides. The GST fusion protein containing BMRF-2-RGD peptide or its mutated form, BMRF-2-AAA, were electrophoresed in Tris-glycine SDS-PAGE gels. Reactivity of human serum samples to these peptides from asymptomatic, NPC and HL groups was examined by Western blot assay. As a negative control EBV-negative human serum was used (Wampole Laboratories). The GST protein was detected as a loading control.

Jianqiao Xiao, et al. Virology. ;393(1):151-159.
4.
Fig. 1

Fig. 1. From: EBV-positive human sera contain antibodies against the EBV BMRF-2 protein.

Expression of BMRF-2 in Akata/BMRF-2 and 293T/BMRF-2 cells. (A) Cells transfected with the BMRF-2-GFP fusion gene were fixed, and the GFP fluorescence signal was visualized in cells using confocal microscopy. Cell nuclei were counterstained in blue. (B) Membrane fractions of Akata/BMRF-2 and 293T/BMRF-2 cells were isolated and 10 μg of total membrane proteins were separated in urea gels. BMRF-2 was detected by Western blot assay using rat anti-BMRF-2 antiserum.

Jianqiao Xiao, et al. Virology. ;393(1):151-159.
5.
Fig. 5

Fig. 5. From: EBV-positive human sera contain antibodies against the EBV BMRF-2 protein.

Neutralizing activity of EBV-positive human sera pre-treated with BMRF-2 peptides. Human serum samples from asymptomatic, NPC and HL patients were incubated with the 47 aa BMRF-2 RGD or BMRF-2 AAA peptides (A and B) for 1 h and then used for neutralization of EBV B95-8 virus. In parallel experiments (C) the serum samples were incubated with a shorter (17 aa) BMRF-2 RGD (RRRSIFCARGDHSVASL) or a control, unrelated peptide (GARRNQIYTSGLERRR). Polarized primary tongue epithelial cells were infected with EBV B95-8 pre-incubated with human serum samples treated with or without peptides. (A) At 3 days post-infection, cells were immunostained using anti-BZLF-1 antibody and examined by confocal microscopy. Only merged panels are shown; yellow indicates colocalization of BZLF-2 (green) signals with the nuclear marker (red). (B, C) EBV-infected (BZLF-1-positive) cells were counted, and the percentage of cells infected was determined for each experimental condition. (B) Asterisks indicate statistically significant inhibition of EBV-neutralizing activity using serum samples treated with BMRF-2 peptides as compared with serum samples without peptides (EBV+ sera) (*P<0.05). (C) Asterisks indicate statistically significant inhibition of EBV-neutralizing activity using serum samples treated with a BMRF-2 peptide as compared with serum samples preincubated with the BMRF-2 unrelated peptide (*P < 0.05). Error bars show ± s.e.m. (n = 6). EBV+ sera, pool of 15 samples of EBV-positive sera from asymptomatic, NPC and HL donors. EBV– sera, pool of 2 EBV-negative sera from commercial sources.

Jianqiao Xiao, et al. Virology. ;393(1):151-159.

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